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The human dental pulp stem cells (hDPSCs) are one of the readily available sources of multipotent mesenchymal stem cells (MSCs) and can be considered as a type of tool cells for cell‐based therapies. However, the main limitation in the clinical use of these cells is DPSC senescence, which can be induced by lipopolysaccharide (LPS) of oral pathogenic bacteria. Up to now, far little attention has been paid to exploring the molecular mechanisms of senescence in DPSCs. So, the current study aimed to investigate the underlying molecular mechanism of senescence in hDPSCs stimulated with Porphyromonas gingivalis (P. gingivalis) and Escherichia coli (E. coli)‐derived LPSs, by evaluating both mRNA and protein expression of four important senescence‐related genes, including TP53, CDKN1A, CDKN2A and SIRT1. To this purpose, hDPSCs were stimulated with different LPSs for 6, 24 and 48 h and then the gene expression was evaluated using quantitative real‐time polymerase chain reaction (qPCR) and western blotting. Following stimulation with P. gingivalis and E. coli‐derived LPSs, the relative mRNA and protein expression of all genes were significantly up‐regulated in a time‐dependent manner, as compared with unstimulated hDPSCs. Moreover, the hDPSCs stimulated with P. gingivalis LPS for 6 and 24 h had the highest mRNA expression of CDKN1A and SIRT1, respectively (p < 0.0001), whereas the highest mRNA expression of CDKN2A and TP53 was seen in hDPSCs stimulated with E. coli LPS for 48 h (p < 0.0001). In summary, because DPSCs have been reported to have therapeutic potential for several cell‐based therapies, targeting molecular mechanisms aiming at preventing DPSC senescence could be considered a valuable strategy. [ABSTRACT FROM AUTHOR]

Introduction: Osteopontin (OPN), plays an important role in immune system modulation. OPN can activate osteoclasts, thus causing resorption of bone. In addition, it might have a protective function against polymicrobial endodontic infections. Since different isoforms of OPN might have diverse roles, the aim of the present study was to compare gene expression of different isoforms of osteopontin in symptomatic irreversible pulpitis and normal pulps of the human dental pulp. Materials and Methods: Pulps were taken from 20 teeth with symptomatic irreversible pulpitis as the case group and from 20 intact premolars scheduled for extraction as the control group. After RNA extraction and synthesis of complementary DNA (cDNA), quantitative real-time polymerase chain reaction (PCR) was used for the evaluation of gene expression of OPN, OPN2 and OPN3. The Mann-Whitney U, t and Chi-square tests were used to analyze differences between the groups. Results: Mean values of OPN, OPN2 and OPN3 in normal pulps were 0.695±0.295, 0.656±0.298 and 0.816±0.422, respectively. Mean values of OPN, OPN2 and OPN3 in symptomatic irreversible pulpitis were 2.52±1.82, 1.99±0.899 and 1.816±0.954, respectively. Unlike OPN and OPN2, OPN3 exhibited significantly higher expression in normal pulps (P<0.05). Conclusion: The results of the present case-control study showed that some variants of OPN are upregulated during pulpitis and it might be due to their prominent modulatory roles in dental pulps. [ABSTRACT FROM AUTHOR]

The progression of periodontitis depends on interactions between the periodontal pathogens and the host immune cytokines, including interleukin (IL)-1β and IL-18. Production of IL-1β is regulated by NOD-like receptors family pyrin domain containing 3 (NLRP3). This study aimed to evaluate the effect of periodontal treatment on the concentrations of IL-18 and NLRP3 in patients with chronic periodontitis. In this experimental study, 18 patients with chronic periodontitis and a mean age of 46.2±8.95 years, were included. The gingival crevicular fluid (GCF) was collected at the beginning of the study, 4 weeks after non-surgical (phase I), and 4 weeks after surgical periodontal treatment. The levels of NLRP3 and IL-18 were measured; using an enzyme-linked immunosorbent assay. Pearson correlation test was used to analyze the concentration of NLRP3 and IL-18 before and after the treatments with CAL and PD. There was a significant association between the level of NLRP3 and the mean values of PD and CAL before treatment. After each treatment phase, a significant decrease was observed in the NLRP3 level. There was no significant relationship between IL-18 and clinical parameters before and after periodontal treatments. Given the possible association between the level of NLRP3 and clinical parameters, we suggest it as a possible indicator of inflammation in chronic periodontitis and an index for evaluating the treatment outcome. [ABSTRACT FROM AUTHOR]

Introduction: Periodontitis is a disease of the periodontal tissues that results in attachment loss and destruction of alveolar bone. Several risk factors such as bacteria, the host immune responses and genetics are considered. Aim: The present study was undertaken to evaluate the effect of phase I periodontal treatment on the concentrations of IL-22 and S100A12 in patients with chronic periodontitis. Correlation between PD and CAL with the concentrations of IL-22 and S100A12 was also evaluated. Materials and Methods: In this study, 22 patients (14 female and 8 male), with a mean age of 39.55±10.25 years, and moderate to severe periodontitis were included. GCF samples were obtained and then Phase I periodontal treatment was rendered. The subjects were recalled after four weeks for collecting samples. Wilcoxon's signed-rank test was used to analyse concentration of S100 and IL-22 before and after the treatment with CAL and PD. Results: There was an inverse correlation between the mean PD and S100 before treatment (p<0.05). In relation to CAL, although an inverse correlation was expected between CAL and S100 concentration, no significant correlation was found between them (p≈0.079). Conclusion: The periodontal tissues are weak cellular sources for the secretion of IL-22 and S100. Given the inverse correlation between the concentration of S100 and the mean PD before treatment and also the inverse and significant correlation between the concentration of S100 and the decrease in CAL and PD after treatment, a possible protective role might be considered for S100 for the incidence of periodontitis and success of phase I treatment. [ABSTRACT FROM AUTHOR]

Introduction: Interleukin-17 is a pro-inflammatory cytokine with a wide range of protective and destructive effects in periodontitis. The role of IL-23 is stabilisation and expansion of Th-17. The aim of this study was to assess whether patients with aggressive and chronic periodontitis exhibit different gingival crevicular fluid (GCF) concentrations of IL-17 and IL-23 compared with clinically healthy subjects. Material and methods: GCF samples were obtained from 32 patients: 10 with chronic periodontitis (CP), 12 with aggressive periodontitis (AgP), and 10 healthy controls (HC). IL-23 and IL-17 concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Comparison of study groups was performed with ANOVA and Tukey HSD tests. Spearman's correlation coefficient was used to assess correlations between the variables. Results: IL-17 concentration was significantly higher in the healthy group compared to the AgP and CP groups (p < 0.001), but there were no significant differences between the CP and AgP groups. IL-23 levels in the healthy group were significantly higher than that in the AgP group (p < 0.001). Cytokine concentrations did not correlate significantly with probing depths and clinical attachment levels. Conclusions: Gingival crevicular fluid concentrations of IL-17 and IL-23 were significantly higher in the healthy group compared to periodontitis groups. [ABSTRACT FROM AUTHOR]

The present study aimed to compare the levels of high-mobility group box 1(HMGB1) and soluble triggering receptor expressed on myeloid cells (sTREM1) in the gingival crevicular fluid (GCF). This cross-sectional cohort trial investigated two groups of 22 eligible chronic periodontitis and 22 periodontally healthy individuals (student volunteers) both before and after the periodontal treatment.  GCF was collected from the deepest pockets with clinical attachment loss≥3 mm. Both groups received oral hygiene instructions, and scaling and root planning were performed in the test group. Enzyme-linked immunosorbent assay kit (ELISA) was used to measure the levels of HMGB1 and sTREM1 in GCF samples collected before and 1 month after non-surgical periodontal treatment. The results showed that HMGB1 levels were significantly higher in the chronic periodontitis patients than those of the healthy individuals before treatment (p<0.02) and decreased significantly after periodontal treatment, which reduced gingival inflammation. Furthermore, the levels of sTREM1 marker were significantly higher in periodontitis patients before (p<0.001) and 1 month after treatment than in healthy individuals (p<0.003) although its crevicular levels decreased after periodontal therapy in periodontitis group. The higher levels of sTREM1 and HMGB1 cytokines in GCF of periodontitis patients and the significant decrease after the introduction of the periodontal treatment underlines the importance of HMGB1 and sTREM1 in pathogenesis of periodontitis. [ABSTRACT FROM AUTHOR]

One of the inflammatory mediators which is secreted by inflammatory cells is high-mobility group protein B1 (HMGB1). Interaction of HMGB1 and toll-like receptors (TLRs) leads to increased production of inflammatory cytokines. On the other hand, it was shown that triggering receptor expressed on myeloid cells (TREM-1) also can be activated by TLRs, and its soluble form (sTREM-1) can be formed by cleaving of membrane-bound form of TREM-1 proteinases. Since there is not enough knowledge about the precise role of HMGB1 and sTREM-1 in periodontal diseases, the aim of this study was to evaluate the concentration of HMGB1 and sTREM-1 in gingival crevicular fluid (GCF) samples of patients with chronic periodontitis. Gingival crevicular fluid (GCF) samples were obtained from a total of 24 individuals with clinically healthy gingiva and 24 patients with moderate to severe chronic periodontitis. For collecting GCF samples, periopapers were placed at the entrance of the crevice and left in position for 30 seconds. Then, they were stored at -80°C. Enzyme-linked immunosorbent assay (ELISA) was used for measuring the concentration of HMGB1 and sTREM-1 in GCF samples. The concentration of HMGB1 (p<0.001) and sTREM-1 (p<0.017), was significantly higher in chronic periodontitis group. In addition, there was a significant positive correlation between HMGB1 and sTREM-1 concentration in chronic periodontitis group (p<0.05). We also found significant positive correlation between PD (Pocket depth) and the concentration of HMGB1 (p<0.001) and sTREM-1 (p<0.015). It is concluded that both HMGB1 and sTREM-1 are released during the inflammatory response of periodontal tissues and they can promote inflammatory process, which leads to tissue destruction. [ABSTRACT FROM AUTHOR]

Background: MicroRNAs (miRNAs) are a group of small non-coding RNAs that play an important role in the regulation of gene expression. miRNA-146a (miR-146a), a member of the miR-146 family, is involved in the control of inflammation. Periodontitis is a set of chronic inflammatory disorders of the tissues surrounding the teeth that lead to the breakdown of alveolar bone and tooth loss. In this study, expression levels of miR-146a and its targets, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, are evaluated in human patients with chronic periodontitis (CP).Methods: The study population consisted of 10 healthy controls and 20 individuals with CP. For each participant, clinical parameters including probing depth and clinical attachment level were measured, and a gingival tissue sample was collected. Levels of miR-146a, TNF-α, IL-1β, and IL-6 were quantified using real-time polymerase chain reaction.Results: Levels of miR-146a were significantly higher in patients with CP (P <0.001). There was a positive correlation between levels of miR-146a and clinical parameters (P <0.05). Elevated miR-146a was accompanied by a significant reduction in TNF-α and IL-6 (P <0.001).Conclusions: Patients with CP had higher levels of miR-146a than healthy individuals, accompanied by reduced levels of TNF-α and IL-6. A positive relationship between miR-146a levels and clinical parameters suggests a pathophysiologic role of miR-146a in CP. [ABSTRACT FROM AUTHOR]

Background and aims. Lysozyme and lactoferrin are salivary proteins which play an important role in innate defense mechanisms against bacteria. This study investigated the association of salivary lysozyme and lactoferrin concentrations with early childhood caries (ECC). Materials and methods. This study was carried out on 42 healthy children (age range, 36 to 71 months), of whom 21 were caries free (CF) and 21 had ECC. Disposable needle-less syringes were used to collect unstimulated saliva from buccal and labial vestibules. Fifteen children who had ECC were treated completely and their saliva was collected in the same way for the second time, three months after treatment. Lysozyme and lactoferrin concentrations were measured and recorded by the ELISA method. The intergroup comparisons were carried out using chi-square, Student's t-test and Wilcoxon signed ranked test. A P-value less than 0.05 was considered as statistically significant. Results. The mean concentration of lysozyme was significantly higher in CF group compared with that of ECC group (P = 0.04). Although the mean concentration of lactoferrin in ECC group was higher in comparison with ECC group, the difference was not statistically significant (P = 0.06). After dental treatment, the mean concentrations of lysozyme and lactoferrin did not change in comparison with their concentrations before treatment. Conclusion. ECC may have a relationship with lower concentrations of unstimulated salivary lactoferrin and lysozyme and reduced amounts of these two salivary proteins may be a risk factor for dental caries in children. [ABSTRACT FROM AUTHOR]

Background: Osteoclastogenesis is coordinated by the interaction of three members of the tumor necrosis factor (TNF) superfamily: Osteoprotegerin (OPG)/receptor activator of nuclear factor kappa B ligand (RANKL)/receptor activator of nuclear factor kappa B (RANK). The aim of this study was to investigate RANKL and OPG levels, and their relative ratio in gingival crevicular fluid (GCF) of patients with chronic and aggressive periodontitis, as well as healthy controls. Materials and Methods: In this analytical study, GCF was obtained from healthy (n = 10), mild chronic periodontitis (n = 18), moderate chronic periodontitis (n = 18), severe chronic periodontitis (n = 20), and generalized aggressive periodontitis (n = 20) subjects. RANKL and OPG concentrations were measured by enzyme-linked immunosorbent assay. Statistical tests used were Kruskal-Wallis test, Mann-Whitney U rank sum test, and Spearman's rank correlation analysis. The level of statistical significance was set at P < 0.05. Results: Mean RANKL concentration showed no statistically significant differences between groups (P = 0.58). There were also no significant differences between mean OPG concentration in the five groups (P = 0.0.56). Moreover, relative RANKL/OPG ratio did not reveal a significant difference between the three study group subjects: healthy, chronic periodontitis (mild, moderate, severe), and aggressive periodontitis (P = 0.41). There was statistically significant correlation between the concentration of sRANKL and Clinical Attachment Level (CAL) in moderate chronic periodontitis patients (R = 0.48, P = 0.04). There was also negative correlation between OPG concentration and CAL in moderate chronic periodontitis patients, although not significant (R = .0.13). Conclusion: RANKL was prominent in periodontitis sites, especially in moderate periodontitis patients, whereas OPG was not detectable in some diseased sites with bleeding on probing, supporting the role of these two molecules in the bone loss developed in this disease. [ABSTRACT FROM AUTHOR]

Objective: Resin cements, regardless of their biocompatibility, have been widely used in restorative dentistry during the recent years. These cements contain hydroxy ethyl methacrylate (HEMA) molecules which are claimed to penetrate into dentinal tubules and may affect dental pulp. Since tooth preparation for metal ceramic restorations involves a large surface of the tooth, cytotoxicity of these cements would be more important in fixed prosthodontic treatments. The purpose of this study was to compare the cytotoxicity of two resin cements (Panavia F2 and Rely X Plus) versus zinc phosphate cement (Harvard) using rat L929-fibroblasts in vitro. Materials and Methods: In this experimental study, ninety hollow glass cylinders (internal diameter 5-mm, height 2-mm) were made and divided into three groups. Each group was filled with one of three experimental cements; Harvard Zinc Phosphate cement, Panavia F2 resin cement and Rely X Plus resin cement. L929-Fibroblast were passaged and subsequently cultured in 6-well plates of 5×105 cells each. The culture medium was RPMI_1640. All samples were incubated in CO2-. Using enzyme-linked immune-sorbent assay (ELISA) and (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) (MTT) assay, the cytotoxicity of the cements was investigated at 1 hour, 24 hours and one week post exposure. Statistical analyses were performed via two-way ANOVA and honestly significant difference (HSD) Tukey tests. Results: This study revealed significant differences between the three cements at the different time intervals. Harvard cement displayed the greatest cytotoxicity at all three intervals. After 1 hour Panavia F2 showed the next greatest cytotoxicity, but after 24-hours and oneweek intervals Rely X Plus showed the next greatest cytotoxicity. The results further showed that cytotoxicity decreased significantly in the Panavia F2 group with time (p<0.005), cytotoxicity increased significantly in the Rely X Plus group with time (p<0.001), and the Harvard cement group failed to showed no noticeable change in cytotoxicity with time. Conclusion: Although this study has limitations, it provides evidence that Harvard zinc phosphate cement is the most cytotoxic product and Panavia F2 appears to be the least cytotoxic cement over time. [ABSTRACT FROM AUTHOR]

The purpose of this study was to compare substance P (SP) and calcitonin gene-related peptide (CGRP) expression in pulp tissue with clinically diagnosed symptomatic and asymptomatic irreversible pulpitis. Healthy pulps acted as controls. Five normal pulps and 40 with irreversible pulpitis (20 symptomatic and 20 asymptomatic) were obtained from 45 different patients. SP and CGRP expression was determined by competition binding assays using enzyme immunoassay.anova and Mann–Whitney tests were used to ascertain if there were statistically significant differences between the groups. The results showed that neuropeptides were found in all pulp samples. The highest and the lowest expressions for SP and CGRP were found in symptomatic irreversible pulpitis and healthy pulps groups, respectively. The differences between healthy pulps and the groups of pulps having irreversible pulpitis were significant ( P < 0.001). Although Mann–Whitney's post-hoc tests showed statistically significant differences in CGRP expression between two pulpitis groups ( P < 0.05), differences in SP expression between symptomatic and asymptomatic irreversible pulpitis groups were not significant. This study demonstrated that the expression of CGRP and SP is significantly higher in pulps with irreversible pulpitis compared with healthy pulps. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the relationship between pulp polyp formation and immunoglobulin E (IgE), histamine and interleukin-4 (IL-4) as the most important mediators which are involved in allergy. Thirty-two samples including 16 pulp polyps and 16 normal pulps were gathered. After homogenising the pulpal tissue samples, enzyme-linked immunosorbent assay (ELISA) techniques were used to assess the concentration of IgE, histamine, IL-4 and IL-12. The two groups showed statistically significant differences in terms of both the concentration and presence of IgE, histamine and IL-4 ( P < 0.001); both presence and concentration of IgE, histamine and IL-4 were higher in pulp polyps than in normal pulps. There is not any significant difference between case and control groups regarding IL-12. The results of this study give rise to the possibility of type I hypersensitivity reaction being involved in pulp polyp's pathogenesis. [ABSTRACT FROM AUTHOR]

Background: Chitin microparticles (CMPs) are found to be potent macrophage stimulators; however, their immunomodulatory effects on the parasite-infected macrophages have not yet been studied. To address this issue, we used a Leishmania major-infected murine macrophage model and characterized the regulatory effects of CMPs on the parasite-infected cells. Methods: Mouse peritoneal macrophages were prepared and infected with L. major (MRHO/IR/1975/ER) standard strain. Following cell treatment with CMPs (500 μg/mL) for 48 h, percent of infected macrophages was determined by Giemsa staining and compared with untreated cells. To find the potential mechanisms of the activity of CMPs, TNF-α and accumulated nitrite in the culture supernatants of the treated and untreated cells were also measured by ELISA and colorimetric Griess assays, respectively. Results: According to the obtained results, chitin microparticles reduced the ex vivo parasite infectivity by ∼12%. However, this inhibitory effect was not directly related to the increased biosynthesis and release of nitric oxide (NO) by macrophages. Instead, we observed a significant increase in the level of TNF-α secretion due to cell treatment with CMPs. Interestingly, this overexpression of TNF-α did not impair cell viability, suggesting the anti-apoptotic effects of the CMPs. Conclusions: These findings show that chitin microparticles have immunomodulatory effects on L. major-infected macrophages and further provide motivations for future studies on their in vivo effects. [ABSTRACT FROM AUTHOR]